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Shim, Sang-Hee
Shim Research Lab
Research Interests
  • Super-resolution Optical Imaging
  • Single-molecule Biophysics
  • Ultrastructural Dynamics
  • Single-cell Systems Biology

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Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes

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Title
Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes
Author
Shim, Sang-HeeXia, ChenglongZhong, GuishengBabcock, Hazen P.Vaughan, Joshua C.Huang, BoWang, XunXu, ChengBi, Guo-QiangZhuang, Xiaowei
Keywords
Diffraction limit; Nanoscopy; Photoactivation localization microscopy; Photoswitchable dye; Stochastic optical reconstruction microscopy
Issue Date
2012-08
Publisher
NATL ACAD SCIENCES
Citation
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.109, no.35, pp.13978 - 13983
Abstract
Imaging membranes in live cells with nanometer-scale resolution promises to reveal ultrastructural dynamics of organelles that are essential for cellular functions. In this work, we identified photoswitchable membrane probes and obtained super-resolution fluorescence images of cellular membranes. We demonstrated the photoswitching capabilities of eight commonly used membrane probes, each specific to the plasma membrane, mitochondria, the endoplasmic recticulum (ER) or lysosomes. These small-molecule probes readily label live cells with high probe densities. Using these probes, we achieved dynamic imaging of specific membrane structures in living cells with 30-60 nm spatial resolution at temporal resolutions down to 1-2 s. Moreover, by using spectrally distinguishable probes, we obtained two-color super-resolution images of mitochondria and the ER. We observed previously obscured details of morphological dynamics of mitochondrial fusion/fission and ER remodeling, as well as heterogeneous membrane diffusivity on neuronal processes.
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DOI
10.1073/pnas.1201882109
ISSN
0027-8424
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BME_Journal Papers
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